Aim: This study aims to develop and to standardize a polymerase chain reaction (PCR) assay that will diagnose clinical as well as carrier state of the disease and to compare the results with conventional microscopy technique.

Materials and Methods: A herd of crossbred cattle with the previous history of theileriosis in village Lahli, district Rohtak, Haryana, was selected for this study. A total of 29 blood samples were collected randomly from cows including five clinically ill cattle. Blood smears from all animals and lymph node biopsy smears from animal with swollen lymph nodes were examined microscopically after conventional Giemsa staining. Phenol chloroform isoamyl alcohol method was used for extracting DNA from blood. Previously published primers targeting cytochrome b gene sequence of Theileria annulata were used in the PCR assay that was standardized to use in the laboratory.

Results: Out of 29 samples tested,18 (62.06%) were found positive for theileriosis by PCR assay, whereas only 10 (34.48%) samples were detected positive by conventional microscopic technique using Giemsa staining method.

Conclusions: On the basis results of comparative studies, it can be concluded that PCR assay is a more sensitive than microscopic examination for detection of theileriosis. This can be attributed to the ability of PCR assay to detect small amounts of genomic DNA of T. annulata or low parasitemia in cows. Therefore, PCR assay can serve as a more sensitive tool to detect Theileria for detection of theileriosis even in asymptomatic carrier cattle which is important for the implementation of successful control programs.

Aim: Fascioliasis is one of the most zoonotic diseases with global extension. As the epidemiological distribution of Fasciola may lead to various genetic patterns of the parasite, the aim of this study is to identify Fasciola hepatica based on spermatogenesis, and phylogenetic analysis using mitochondrial (nicotiamide adenine dinucleotide dehydrogenase subunit I [ND1] and cytochrome oxidase subunit I) gene marker.

Materials and Methods: In this study, 90 F. hepatica collected from 30 cattle at slaughterhouse located in three different geographical locations in the North-East of Iran were evaluated based on spermatogenetic ability and internal transcribed spacer 1 gene restriction fragment length polymorphism pattern. Genetic diversity and phylogenetic relationship using mtDNA gene marker for the isolates from the North-East of Iran, and other countries were then analyzed.

Results: Partial sequences of mtDNA showed eight haplotypes in both genes. The phylogenic analysis using neighbor joining as well as maximum likelihood methods showed similar topologies of trees. Pairwise fixation index between different F. hepatica populations calculated from the nucleotide data set of ND1 gene are statistically significant and show the genetic difference.

Conclusion: F. hepatica found in this region of Iran has different genetic structures through the other Fasciola populations in the world.

Aim: The objective of this study was to screen the prevalence of contagious ecthyma (CE) among the goat population of Assam owing to its high prevalence rate.

Materials and Methods: In this study, a total of 231 serum samples were collected from 12 districts of Assam during September 2013 to July 2014. The serum samples were tested for the presence of antibodies against Orf virus (ORFV) by indirect enzyme-linked immunosorbent assay (ELISA). Indirect ELISA was standardized using purified Orf reference virus produced in bulk in primary lamb testes cells.

Results: Studies on seroprevalence showed 76.62% of goats were seropositive. The total number of animals were divided into different age groups starting from 0-2 months, 2-4 months, 4-6 months, and above 8 months and accordingly highest prevalence of antibodies against ORFV was recorded in the age-group above 8 months of age. Significantly, lower rates of infection were observed in goats of age group 2-4 months. This study recorded that seropositivity from naturally infected animals and in contact apparently healthy animals to be 53.67% and 46.32%, respectively.

Conclusion: The results indicated that CE is a prevalent infection in goats of Assam, and the healthy population is at increased risk of infection.

Aim: We describe a laboratory investigation carried out to confirm the etiology of the heavy mortality (37 animals died out of total 44, i.e. 84%) in goats in Ri-Bhoi district of Meghalaya, Northeast region of India in December 2015. The clinical signs observed were abortion, diarrhea, high fever (up to 104°F), pox lesion in the skin, and respiratory distress.

Materials and Methods: The samples comprising whole blood, sera, and pox lesion were collected from the animals (n=7) from an outbreak for the screening of peste des petits ruminants (PPR) and poxviruses. The whole blood and sera were used for screening of PPR virus (PPRV) by sandwich enzyme-linked immunosorbent assay (ELISA) and antibody by competitive ELISA as well as detection of PPRV partial N gene by reverse transcription-polymerase chain reaction (PCR). The skin lesions were used for the detection of poxvirus by PCR.

Results: The results showed the presence of PPR antigens (58-80%) in the samples by sandwich ELISA and antibody in all the sera samples ranging from 9% to 41% positivity in competitive ELISA. Four samples were positive for PPRV partial N gene. The skin lesion screened for poxvirus was also found to be positive for I3L gene of goatpox virus.

Conclusion: We confirm the outbreak of disease in goats with high mortality is a case of mixed infection of PPR and goatpox detected for the first time in Northeast India.

Brucellosis is the most common worldwide zoonosis with 500,000 new cases every year in humans and infections in millions of animals. This infection is mainly acquired by humans through consumption of unpasteurized milk and milk products from infected animals. Exposure can also occur occupationally in those who work closely with animals through contact with aborted fetuses and reproductive secretions. Animals acquire the infection from other infected animals through direct contact and vertical transmission. This infection is prevalent in all continents of the world except Antarctica, but its impact is more felt in developing countries where it is endemic in animals and humans. In certain developed countries where the disease was eradicated, there seem to be a re-emergence of the disease as the disease appears to claim more territory. The risk factors of the disease may vary from country to country and region to region, but most risk factors are similar. Consumption of unpasteurized milk and milk products plays a very important role in the transmission of this infection from animals to humans, in addition to direct contact with infected animals and their secretions. The best way to control this ubiquitous infection is through the One Health approach which involves human health, animal health, and environmental health. This paper reviews the prevalence of brucellosis in some countries in various continents of the world and highlights the risk factors responsible for the persistence of this infection in animals and humans with a view to proffering solution to this age-old zoonosis that has defied eradication for many generations in many parts of the world.

Aim: This study was conducted to assess the extent of knowledge, awareness, attitude, and risks of zoonotic diseases among livestock owners in Puducherry region.

Materials and Methods: A total of 250 livestock farmers were selected randomly from eight revenue villages. And each farmer was interviewed with a questionnaire containing both open- and close-ended questions on various aspects of zoonotic diseases, a total of 49 questionnaires were framed to assess the source and transmission of infection to the farmers and to test their knowledge and awareness about zoonotic diseases. The data collected were analyzed by chi-square test using software Graph pad prism, and results were used to assess the relationship between education level and zoonotic disease awareness; risk of zoonotic diseases and its relation with independent variables.

Results: The present survey analysis represents that most of the respondents are belonging to the age group of 41-60 years. About 42.8% of respondents’ household having a graduate. The most of the respondent are small-scale farmers and their monthly income was less than Rs. 10,000. About 61.2% of farmers were keeping their animal shed clean. About 29.6% of the respondents were ignorant about cleaning the dog bitten wound. Only 16.4% of respondents knew that diseases in animals can be transmitted to humans. Only 4.8%, 3.6%, 6.8%, and 22.4% of respondents knew about the zoonotic potential of diseases such as brucellosis, tuberculosis (TB), anthrax, and avian flu, respectively. Only 18% of the respondents were aware about zoonotic diseases from cattle. Regarding the list of zoonotic diseases contracted, 37.7% reported respiratory infection, 31.1% digestive disturbances, 15.5% had dermatological problem, and 15.5% reported indiscrete disease such as fever, body pain, and headache joint pain. From the respondent got the zoonotic disease (n=45), 51.2% of the respondent reported chronic infection and 48.8% of the respondent reported acute form of zoonotic infection. About 30% of the respondents’ farm had an incidence of abortion. Our analyses showed that there was significant in educational level of respondents and treatment of dog bitten animals. Furthermore, there was statistical significance in occurrence of hand and foot lesions in the respondent and occurrence of foot-and-mouth disease outbreak in their animals.

Conclusion: From this study, it is concluded that involvement of educated family members in farming practices can create awareness and improve knowledge toward zoonotic disease. Further creation of awareness toward zoonotic diseases is of utmost important.

Aim: The present investigation was undertaken to study the reproduction performance and effect of non-genetic factors on reproduction performance of Jersey crossbred cows.

Materials and Methods: Data on 355 Jersey crossbred cattle maintained at the Post-graduate Research Institute in Animal Sciences, Kattupakkam, Tamil Nadu, distributed over 30 years (1985 to 2014). The effect of various non-genetic factors including the period of birth, season of birth, period of calving, season of calving and parity were analyzed through least-squares analyses using univariate general linear model. The different (co)variance components for calculation of genetic parameters were estimated using restricted maximum likelihood method by fitting an animal model.

Results: The overall least-squares means (±standard error) of age at first service, age at first calving, weight at first calving, service period, calving interval, dry period, and number of services per conception were 848.06±9.72 days, 1204±12.20 days, 289.81±1.71 kg, 210.01±6.41 days, 489.12±6.45 days, 137.96±5.58 days, 2.50±0.07, respectively. Period of calving had either significant (p<0.05) or highly significant (p<0.01) effect on all reproduction traits studied except service period, calving interval, and dry period. Number of services per conception was affected by season of calving. Parity had significant influence (p<0.05) or highly significant (p<0.01) influence on all the traits studied except service period and dry period. Heritability estimates of age at first service, age at first calving, weight at first calving, service period, calving interval, dry period, and number of services per conception were 0.299, 0.220, 0.017, 0.142, 0.222, 0.177, and 0.042, respectively. The estimates of repeatability for service period, calving interval, dry period, and number of services per conception were 0.219, 0.234, 0.420, and 0.001, respectively.

Conclusions: The reproduction performances of Jersey × Red Sindhi crossbreds were lower when compared to the earlier reports on Jersey crossbreds. Heritability and repeatability values were also low to moderate, indicating limited scope for improvement through selection.

Friday, 23 September 2016

15.Enzyme histochemistry of cecal lymphoid tissue during prenatal period of buffalo -Kritima Kapoor and Opinder Singh

Veterinary World, 9(9): 1006-1011

doi: 10.14202/vetworld.2016.1006-1011

Abstract

Aim: This study was designed to elucidate the histoenzymic distribution of enzymes, i.e., phosphatases, oxidoreductases, dehydrogenases, and diaphorases in cecal lymphoid tissue during its development in the prenatal period.

Materials and Methods: The study was conducted on cecum of 15 buffalo fetuses ranging from 16 cm curved crownrump length (CVRL) (100 days) to 100 cm CVRL (full term). The fetuses were categorized into three groups based on their CVRL.

Results: In Group I, the distribution of enzymes was uniformly weak in developing villi-like projections in cecum and completely absent from submucosa. In Group II, the enzymes showed a moderate to strong activity in epithelium lining tunica mucosa which progressively decreased as the fetus progresses toward late gestational age. However, the intense activity of enzymes was observed in developing lymphoid tissue in this group. In Group III, distribution of enzymes reduced in tunica mucosa of cecum with advancing age, whereas the intense activity was noticed in the developed lymphoid tissue complex.

Conclusion: The distribution of enzymes was completely absent from submucosal region in cecum of Group I as there was no lymphoid tissue development at this age. In Group II, the enzymes showed a moderate to strong activity in epithelium lining tunica mucosa which progressively decreased toward late gestational age but an intense activity was observed in developing lymphoid tissue. In Group III, distribution of enzymes reduced in tunica mucosa with advancing age with intense activity noticed in the developed lymphoid tissue complex.

Materials and Methods: This study examined the expression of CREM on 20 male rats (Rattus norvegicus) at 4 months of age, weighing 250-300 g. The rats were divided into four groups: K0, KP1, KP2, and KP3. K0 group was injected with 0.2 ml normal saline; KP1 was injected with 25 mg cloprostenol (Prostavet C, Virbac S. A); KP2 and KP3 were injected with 0.2 and 0.4 ml seminal vesicle extract, respectively. The treatments were conducted 5 times within 12-day interval. At the end of the study, the rats were euthanized by cervical dislocation; then, the testicles were necropsied and processed for histology observation using immunohistochemistry staining.

Results: CREM expression in rat Sertoli cells was not altered by the administration of either 0.2 or 0.4 ml seminal vesicle extract.

Materials and Methods: A total of 330 samples, comprising 98 chicken meat, 82 chevon meat, 90 raw milk, and 60 human urine and stool samples, were processed for isolation of E. coli. Isolates were confirmed biochemically and further tested against commonly used antibiotics to know their resistant pattern. The resistant isolates were tested for ESBL production by phenotypic method followed by characterization with molecular method using multiplex-polymerase chain reaction technique.

Conclusion: The present study indicates a high prevalence of E. coli in raw chicken meat, chevon meat, and milk due to poor hygienic practices. The antibiotic susceptibility test detected the presence of the resistance pattern against ESBL in E. coli isolated from raw chicken meat, chevon meat, milk, and also in human clinical samples is of great concern. The appearance of E. coli in the human food chain is alarming and requires adaptation of hygienic practices and stipulate use of antibiotics.

Aim: Infectious porcine bronchopneumonia, caused by Pasteurella multocida, is a widespread disease of major economic significance. Thus, the aim of the present study was to diagnose swine Pasteurellosis using gross, histopathological, and immunopathological approaches in the swine population of Punjab and to compare the efficacy of immunohistochemical (IHC) techniques with conventional diagnostic techniques.

Materials and Methods: A total of 71 adult swine lung samples showing gross pneumonic changes were collected along with the associated lymph nodes to carry out the study. The collected samples were then processed for histopathological and IHC studies.

Results: Out of the total 71 lung samples, 26 samples were found to be suspected for Pasteurellosis as per the microscopic changes observed, and out of these 26 samples, 16 cases were confirmed to be positive for Pasteurellosis by IHC. Varied macroscopic changes noted in lungs were pneumonic patches with consolidation of many lobes, congestion, and focal hemorrhages. Main lesions associated with lymph nodes were its enlargement and hemorrhages. Histologically, the lung showed fibrinous and suppurative bronchopneumonia, multifocal suppuration, thickening of septa with fibrin combined with cellular infiltration and edema. The higher IHC expression of P. multocida was seen in the bronchial epithelium besides in alveolar and bronchial exudate. Moreover, on comparing the histopathological and IHC scores which were calculated on the basis of characteristic microscopic lesions and number of antigen positive cells, respectively, a significant positive correlation (r=0.4234) was found.

Conclusion: It was concluded that swine population of Punjab is having P. multocida infection. The gross and histopathological lesions can be helpful in the preliminary diagnosis of Pasteurellosis but needs to be supplemented by other immunodiagnostic tests. Moreover, IHC technique proved to be a specific, reliable, precise, and rapid technique to supplement these conventional methods of diagnosis for Pasteurellosis.

Materials and Methods: In this study, efforts have been made to investigate the effects of three combinations (10+10, 20+20 and 30+30 μg/ml) of PGN and LTA obtained from S. aureus. These antigens were used to challenge the bovine PBMCs. After 6 h of incubation quantitative, real time-polymerase chain reaction was used to study toll-like receptor 2 (TLR-2) and major cytokine mRNA expression in bovine PBMC challenged with three different antigen blends.

Results: The results indicated that mRNA level of interferon gamma is influenced by the expression of TLR-2 gene. Tumor necrosis factor-alpha (TNF-a), interleukin 10 (IL-10), and IL-8 genes showed a maximum response at a dose of 10 μg of PGN and 10 μg of LTA challenge per ml of culture medium. The outcome also suggests that both IL-10 and IL-8 followed the expression pattern of TNF-a.

Conclusion: A dose of 10 μg of PGN and 10 μg of LTA per ml of culture medium was found to be most suitable for challenging PBMC.

Aim: This study investigated the prevalence and antibiotic resistance of Salmonella species isolated from drinking water sources in Tamale Metropolis.

Materials and Methods: Isolation of Salmonella species from 275 different drinking water samples (25 each from dam, well, rain, and bottle, 35 from tap, 40 from water trough, and 100 from sachet) was done using a slightly modified method of the Bacteriological Analytical Manual of the Food and Drugs Administration, USA. 34 Salmonella species isolated from the water samples were examined for their susceptibility to nine different antibiotics using the disc diffusion method. The study was carried out from July 2014 to January 2015.

Results: The overall prevalence of Salmonella species was 4.36% (12/275). Dam 16.00% (4/25) and well 16.00% (4/25) water samples were the most contaminated source, followed by rain water (stored) 12.00% (3/25) and tap water samples 2.86% (1/35). There were no significant differences among water samples which were positive for Salmonella species (p>0.05); however, dam and well samples that were positive forSalmonella species differ significantly (p<0.05) from bottle water, sachet water, and water trough samples, which were negative for Salmonella species. The 34 Salmonella isolates were highly resistant to erythromycin (E) (100%) and vancomycin (VA) (94.12%). Few isolates exhibited intermediate resistances to ceftriaxone (CRO) (17.65%), gentamicin (CN) (17.65%), tetracycline (14.71%), chloramphenicol (C) (5.88%), ciprofloxacin (CIP) (2.94%), and amoxicillin (AMC) (2.94%). Salmonella isolates also exhibited six different antibiotic resistant patterns (VA-E, VA-E-AMC, VA-E-CRO, VA-E-C, VA-E-CRO-AMC, and VA-E-AMC-CN). The resistant pattern VA-E (with multiple antibiotic resistance index of 0.22) was the commonest.

Conclusion: This study indicated that some drinking water sources for humans and animals in Tamale Metropolis are contaminated with Salmonella species which exhibited varying resistance to various antibiotics. Therefore, consumers of water at the Tamale Metropolis are at risk of Salmonella infection from drinking water from positive water sources in the Tamale Metropolis.

10.Cardiac biomarkers and ultrasonography as tools in prediction and diagnosis of traumatic pericarditis in Egyptian buffaloes -Noura E. Attia

Veterinary World, 9(9): 976-982

doi: 10.14202/vetworld.2016.976-982

Abstract

Aim: This study was designed to evaluate the cardiac biomarkers and ultrasonography in prediction and early diagnosis of traumatic pericarditis (TP) in Egyptian buffaloes.

Materials and Methods: A total number of 47 buffaloes were included in the study and divided into two groups: Healthy (n=10) and diseased groups (n=37). Diseased buffaloes were admitted to the Veterinary Teaching Hospital at Zagazig University, Egypt, with a history of anorexia, sudden, and severe reduction of milk production with no response to a previous medical treatment some animals had edema at the dewlap and congestion of the jugulars. These animals were subjected to clinical examination, evaluation by hemato-biochemical analysis including cardiac biomarkers and sonography.

Results: The hemato-biochemical analysis revealed leukocytosis with a shift to left and hyperfibrinogenemia (indicating inflammation). Serum cardiac biomarkers including cardiac troponin I (cTnI), cTnT, nitric oxide, creatine kinase myocardial band, and lactic dehydrogenase enzyme were significantly increased in buffaloes with TP compared with control ones. Ultrasonographically, there were hypoechoic materials with echogenic fibrin interspersed in between the pericardial sac.

Conclusions: The cardiac biomarkers may be considered a useful index in the early diagnosis of TP. Moreover, ultrasonography is an excellent tool for early prediction and diagnosis of such condition.